Understanding the Function of Tamm-Horsfall Protein in Acute kidney Injury.

急性肾损伤（阿基）是住院患者死亡率和发病率的主要决定因素。 了解阿基的病理生理学对于开发治疗至关重要。本申请 提出研究Tamm-Horsfall蛋白（THP）在阿基中的作用。这种蛋白质只表达在 肾由Henle氏粗升支（TAL）细胞构成。在过去的几年里，我们发现了 THP介导阿基中的保护性肾小管串扰。我们发现THP调节炎症信号 和损伤S3段，其是肾脏外髓质中TAL的邻近小管。立即 目前建议的目标是研究THP与S3片段的相互作用在调节 中性粒细胞浸润和限制性氧化应激。这些研究将成为过渡到 治疗应用。事实上，PI的长期目标是了解THP的作用 调节以治疗发生阿基的患者。 中心假设是Tamm-Horsfall蛋白调节由炎症因子引起的炎症反应。 外髓质缺血性损伤;需要THP与周围肾小管相互作用的效应 元素这一假设是根据强有力的初步和已发表的数据提出的。的 以下具体目标将用于调查这一假设。 目的1将研究THP作为阿基后肾脏中性粒细胞浸润的关键调节剂的作用。 目的2研究THP与肾脏相邻上皮细胞相互作用的功能意义 损伤 目的2将研究THP抑制中性粒细胞流入和促进阿基后恢复的治疗调节 这项研究将涉及使用THP敲除和野生型小鼠。用于阿基的动物模型是肾 通过肾蒂夹闭实现缺血再灌注损伤。体外研究将涉及细胞系， 近端小管在这项工作中使用的其他技术包括：免疫荧光激光显微解剖 免疫荧光共聚焦显微镜和活体显微镜，流式细胞术，真实的时间- PCR、蛋白质印迹、各种形式的色谱和质谱。 这项研究是创新的，因为它将揭示THP的新的调节功能， 不仅在肾损伤中，而且在其它形式的急性或慢性肾病中。未来的战略， THP在肾脏中的作用将具有重要的治疗意义。

Acute kidney injury (AKI) is a major determinant of mortality and morbidity in hospitalized patients. Understanding the pathophysiology of AKI is essential for the development of therapy. This application proposes to study the role of Tamm-Horsfall protein (THP) in AKI. This protein is expressed exclusively in the kidney by cells of the thick ascending limbs (TAL) of Henle. In the last few years, we uncovered a key role of THP in mediating a protective tubular cross-talk in AKI. We showed that THP regulates inflammatory signaling and injury to S3 segments, which are neighboring tubules to TAL in the kidney outer medulla. The immediate goal of the current proposal is to study the importance of the interaction of THP with S3 segments in regulating neutrophil infiltration and limiting oxidative stress in AKI. These studies will form the basis to transition into therapeutic applications. Indeed, the PI's long term goal is to understand how the role of THP can be modulated to treat patients who develop AKI. The central hypothesis is that Tamm-Horsfall protein regulates the inflammatory response triggered by ischemic injury in the outer medulla; an effect that requires an interaction of THP with the surrounding tubular elements. This hypothesis has been formulated on the basis of strong preliminary and published data. The following specific aims will be used to investigate this hypothesis. Aim 1 will investigate the role of THP as a key regulator of neutrophil infiltration in the kidney after AKI. Aim 2 will study the functional significance of the interaction of THP with neighboring epithelium in kidney injury Aim 2 will investigate therapeutic modulation of THP to inhibit neutrophil influx and promote recovery after AKI This research will involve the use of THP knockout and wild type mice. The animal model used for AKI is renal ischemia-reperfusion injury achieved through renal pedicle clamping. In vitro studies will involve cell lines for proximal tubules. Other techniques used in this work include: Immunofluorescence laser micro-dissection of specific tubular sections, Immunofluoresence confocal and intravital microscopy, flow cytometry, real time- PCR, western blot, various forms of chromatography and mass spectrometry. This research is innovative, because it will uncover novel regulatory functions for THP that may be relevant not only in kidney injury but also in other forms of acute or chronic renal disease. Future strategies that enhance the role of THP in the kidney will be of important therapeutic significance.